Efficient reproduction of cynomolgus monkey using pronuclear embryo transfer technique

doi:10.1073/pnas.0805639105

Efficient reproduction of cynomolgus monkey using pronuclear embryo transfer technique

*Yunnan Banna Primate Disease Model Research Center, Shanghai Institute of Brain Functional Genomics, Key Lab of Brain Functional Genomics, Ministry of Education and Science and Technology Commission of Shanghai Municipality, East China Normal University, Shanghai 200062, People's Republic of China; and
^‡Brain and Behavior Discovery Institute and Department of Neurology, Medical College of Georgia, Augusta, GA 30912

Communicated by Leon E. Rosenberg, Princeton University, Princeton, NJ, June 10, 2008 (received for review April 2, 2008)

Abstract

One of the technical bottlenecks in producing nonhuman primate models is that current assisted reproductive techniques, such as in vitro culture and frozen conservation of multicell-stage embryos, often result in poor embryo quality and subsequently lead to low birth rates. We investigated whether pronuclear embryo transfer can be used as an effective means for improving pregnancy and live birth rates of nonhuman primates. We collected 174 metaphase II oocytes by laparoscopy from 22 superovulated mature females and then fertilized these eggs using either in vitro fertilization or intracytoplasmic sperm injection, resulting in a 33.3% and a 50% fertilization rate, respectively. These 66 fertilized pronuclear-stage embryos were then tubally transferred to 30 recipients and led to 7 births and 1 abortion. Importantly, we observed that the highest live birth rate of ≈64% was obtained when the transfer of pronuclear embryos was performed in the presence of new corpus luteum in the ovary of recipients between 24 h and 36 h after estradiol peak. Therefore, our experiments demonstrate that by matching the critical time window in the recipient's reproductive cycle for achieving optimal embryo-uterine synchrony, pronuclear embryo transfer technology can significantly improve the pregnancy rate and live birth of healthy baby monkeys. This efficient method should be valuable to the systematic efforts in construction of various transgenic primate disease models.

Footnotes

^†To whom correspondence may be addressed. E-mail: qsun{at}brain.ecnu.edu.cn or jtsien{at}mcg.edu
Author contributions: Q.S. and J.Z.T. designed research; Q.S., J.D., W.Y., Y.J., M.Y., Y.W., and J.Z.T. performed research; P.L.W. and Y.H. contributed new reagents/analytic tools; Q.S. and J.Z.T. analyzed data; and Q.S. and J.Z.T. wrote the paper.
The authors declare no conflict of interest.